Compositions and Methods Related to Calcitriol

ABSTRACT

Contemplated compositions and methods are drawn to use of various boron-containing compounds to temporarily and transiently increase endogenous blood calcitriol concentration. The boron-containing compound is preferably a carbohydrate-boron complex having sufficient stability to achieve measurable quantities of the complex in blood upon oral administration of the complex.

This application claims priority to our copending U.S. provisionalapplication Ser. No. 61/166510 which was filed Apr. 3, 2009, and whichis incorporated by reference herein.

FIELD OF THE INVENTION

The field of the invention is compositions and methods related tomodulation of calcitriol (1,25-dihydroxyvitamin D₃) concentration inblood/serum of a mammal, and especially to short-term modulations ofendogenous calcitriol in human serum.

BACKGROUND OF THE INVENTION

The vitamin D₃ endocrine system, besides playing pivotal roles incalcium homeostasis and bone mineral metabolism, is recognized to play apivotal role in a wide range of fundamental biological functions, andespecially in cell differentiation, inhibition of cell growth, andimmune function. Vitamin D₃ (cholecalciferol) is considered aprohormone, which is converted into the more active form25-hydroxyvitamin D₃ (calcidiol), which in turn is converted into thehormonal form 1,25-hydroxyvitamin D₃ (calcitriol) in selected tissues.Calcitriol then binds and activates the cellular Vitamin D₃ receptor(VDR) to modulate gene transcription and regulate mineral ionhomeostasis. Abrogation of gene activation by VDR action is thought tobe under the control of the calcitriol metabolizing enzyme24-hydroxylase that inactivates calcitriol.

As the VDR receptor (and various isoforms) and the correspondingmetabolic enzymes are expressed in many tissues, it is assumed thatcalcitriol can act in an autocrine, paracrine, or intracrine fashion toaffect the biology of non-classical target tissues. For example,production of both the activating and the metabolizing enzymes by cellsof the immune system suggests that calcitriol can be locally produced inimmune reaction sites. Indeed, recent studies have shown moderateimmunosuppressive effect of calcitriol. Other studies have shown thatcalcitriol may act as a potent antiproliferative agent. For example,Vitamin D deficiency is suspected to raise the risk of prostate cancer,most likely via the VDR but possibly also through its1-alpha-hydroxylation in the prostate. Similarly, calcitriol is acoordinate regulator of proliferation, differentiation, and survival ofbreast cancer cells. In addition, epidemiologic, clinical, and animalstudies suggested that vitamin D status is important for protectionagainst the development of breast cancer. Therefore, vitamin D compoundsthat bind and activate VDRs offer promise as therapeutic agents for thetreatment of established breast cancer.

To increase levels of calcitriol, various attempts have been undertaken.Most commonly, dietary supplementation using Vitamin D₃(cholecalciferol) is considered a viable option to raise cholecalciferollevels with the expectation that an upstream surplus of substrate willultimately lead to the production of the calcitriol form. Unfortunately,calcitriol synthesis and degradation are tightly regulated processes,and no significant increase in calcitriol can be achieved usingadministration of cholecalciferol. Similarly, U.S. Pat. App. No.2005/0032743 teaches use of low-dose administration of selectedboron-containing compounds together with cholecalciferol to raise levelsof calcidiol (25-hydroxyvitamin D₃) in human, presumably via inhibitionof the conversion of the calcidiol to calcitriol. It should be notedthat in this publication, calcidiol (but not calcitriol) was describedas the ‘active form’ of vitamin D₃. In still other attempts, calcitriolis directly administered (ROCALTROL™; 1,25-dihydroxycholecalciferol,commercially available by Roche) to increase calcitriol serumconcentration. However, oral administration of calcitriol is associatedwith a plethora of undesirable side effects, including hypercalcemia,excessive thirst and sweating, nausea, and constipation.

Thus, while numerous compositions and methods of modulating calcitriolconcentrations in serum are known in the art, all of almost all of themsuffer from one or more disadvantages. Consequently, there is still aneed to provide improved compositions and methods to modulate theconcentration of calcitriol in a mammal, and especially in a human.

SUMMARY OF THE INVENTION

The present invention is directed to various compositions and methods oftemporarily and transiently increasing blood calcitriol concentrationusing effective dosages of boron-containing compounds. Most preferably,such compounds are certain carbohydrate-boron complexes with sufficientstability to allow transit into the blood stream upon oraladministration.

In one aspect of the inventive subject matter, a method of method ofacutely and transiently increasing blood calcitriol concentration in amammal includes a step of identifying a dosage and schedule foradministration of a boron-containing compound that is effective toproduce a temporary spike in blood calcitriol concentration in themammal, and another step of providing the boron-containing compound in aformulation that allows administration of the boron-containing compoundin the dosage and schedule effective to produce the temporary spike inthe calcitriol concentration in the mammal.

It is generally preferred that the administration is oraladministration, most preferably at a daily dosage between 100 mg and1000 mg of the boron-containing compound. It is further generallypreferred that the boron-containing compound is a carbohydrate-boroncomplex having a boron portion and at least one carbohydrate ligand(e.g., fructose, mannose, mannitol, sorbose, and/or sorbitol) complexedto the boron portion, typically having a boron-ligand associationconstant of between 3,000 and 20,000. In especially preferred aspects,the boron-ligand association constant that is sufficient to deliver thecarbohydrate-boron complex into blood in measurable quantities. Wherethe complex is electrically charged, it is typically preferred that thecharge is neutralized by a alkaline or earth alkaline cation. Forexample, suitable boron containing complexes include a calcium salt offructoborate. In further contemplated aspects, it is preferred that theboron-containing compound is formulated such that the temporary spike ismeasurable between 30 and 180, and more preferably between 60 and 140minutes post administration.

While not limiting to the inventive subject matter, it is contemplatedthat the temporary spike (e.g., an increase of at least 5% of bloodcalcitriol concentration, and more typically at least of at least 25% ofblood calcitriol concentration) in the blood calcitriol concentration isproduced by activation of blood cells, and especially by activation ofmonocytes. Therefore, and among other suitable uses, it should beappreciated that contemplated compounds may be effective to modulate animmune response, and especially to reduce an inflammatory reaction.

Therefore, and viewed from a different perspective, a method ofmodulating a condition associated with a serum calcitriol concentrationin a mammal (e.g., an inflammatory reaction or a neoplastic condition)is contemplated that includes the steps of identifying a dosage andschedule for administration of a boron-containing compound that iseffective to produce a temporary spike in blood calcitriol concentrationin the mammal; and another step of administering the boron-containingcompound at the dosage and schedule effective to produce the temporaryspike in the calcitriol concentration in the mammal, wherein thetemporary spike is effective to modulate the condition.

As before, it is generally preferred that the administration is oraladministration, and that the dosage is a daily dosage between 100 mg and1000 mg of the boron-containing compound. Most preferably, theboron-containing compound is a carbohydrate-boron complex having a boronportion and at least one carbohydrate ligand complexed to the boronportion, typically having a boron-ligand association constant of between3,000 and 20,000. Consequently, it is also preferred thecarbohydrate-boron complex has a boron-ligand association constant thatis sufficient to deliver the carbohydrate-boron complex into blood inmeasurable quantities.

Various objects, features, aspects and advantages of the presentinvention will become more apparent from the following detaileddescription of preferred embodiments of the invention.

BRIEF DESCRIPTION OF THE DRAWING

FIGS. 1A-1D are exemplary graphs illustrating changes of calcitriollevels in blood of human volunteers 30 minutes (1A), 60 minutes (1B), 90minutes (1C), and 90-120 minutes (1D) after administration ofcontemplated compounds.

FIGS. 2A-2B are exemplary graphs depicting acute increase of bloodcalcitriol over time as a function of administration of contemplatedcompounds.

FIG. 3 is an exemplary graph depicting kinetics of blood calcitriol as afunction of administration of contemplated compounds.

DETAILED DESCRIPTION

The inventor has unexpectedly discovered that various boron-containingcompounds are effective in transiently and acutely raising calcitriollevels in blood of a mammal to a significant degree. Indeed, calcitriollevels could be acutely increased in an amount of at about 80% upon oraladministration of a single dose of a boron-containing complex.

In especially preferred aspects of the inventive subject matter,numerous compositions and methods of modulation of calcitriol areprovided. Most preferably, the modulation is an acute increase in serumconcentration and within a relatively short period of time (e.g., lessthan 3 hours, and more typically within 90-120 minutes), and is oftransient nature (typically lasting less than 12 hours, more typicallyless than 6 hours, and most typically between 30 and 240 minutes). Mosttypically, the acute increase is observable as a temporary spike of thecalcitriol concentration in the blood/serum, wherein the spike istypically an increase of at least 10% over initial/baselineconcentration (more typically at least 20%, even more typically at least50%, and most typically at least 90%) and limited in duration (e.g.,less than 12 hours, more typically less than 8 hours, and most typicallyless than 4 hours).

Thus, it should be appreciated that contemplated compounds typicallyproduce a transient increase in calcitriol blood/serum concentrationrather than a prolonged steady-state increase. Moreover, it should beappreciated that the increase is highly specific towards calcitriol anddoes not significantly affect 25-hydroxyvitamin D₃ (calcidiol). Amongother suitable compounds, it is especially preferred that contemplatedcompounds are boron-ligand complexes, and particularlyboron-carbohydrate and/or boron-amino acid complexes. It should be notedthat the term “boron-containing compound” as used herein expresslyexcludes boric acid and any salt thereof

Therefore, preferred compositions for modulation of calcitriol include aboron-containing compound in which boron or borate is bound (covalentlyor non-covalently) in a complexed form with at least one ligand, andmore typically between two and four ligands. For example, suitableligands will include those in which at least one of the ligands includesoxygen, nitrogen, carbon, and/or sulfur, and even more preferably inwhich all of the ligands include at least one of the above atoms. Viewedfrom a different perspective, ligands are especially preferred where theligand(s) provide a relatively high association constant, mostpreferably in the range of about 3,000 to about 20,000.

Among other advantages, it is contemplated that boron complexes with arelatively high association constant will have sufficient stability invivo to survive the passage through the gastrointestinal system andabsorption in to the blood stream in an unaltered form. While notlimiting to the inventive subject matter, the inventor hypothesizes thatstable boron-containing complexes will interact with certain mammaliancells (and especially monocytes and proximal tubule cells of thenephron) to stimulate 1-alpha hydroxylase, directly or indirectly(possibly via parathyroid hormone receptor PTH1R and/or PTH2R).

Consequently, suitable ligands in contemplated complexes includesaccharides (mono-, di-, and poly-) natural and synthetic amino acids,polyols, etc. Thus, especially preferred ligands have a conformationwith at least two hydroxyl groups, or one hydroxyl group and one aminogroup in a 1,2- and a 1,3- position relative to each other. For example,suitable ligands include fructose, sorbitol, mannitol, xylitol, sorbose,serine, threonine, etc. Additionally contemplated boron-containingcompounds are described in U.S. Pat. Nos. 5,962,049, 5,985,842, and6,080,425, and U.S. Pat. App. No. 2005/0032743, all of which areincorporated by reference herein. This and all other extrinsic materialsdiscussed herein are incorporated by reference in their entirety. Wherea definition or use of a term in an incorporated reference isinconsistent or contrary to the definition of that term provided herein,the definition of that term provided herein applies and the definitionof that term in the reference does not apply.

While synthetic compounds are preferred in at least some aspects of theinventive subject matter, it should be recognized that allnaturally-occurring boron/borate-containing compounds are also deemedsuitable for use herein. Especially suitable compounds may be isolatedfrom plant materials (and particularly from cell wall associatedmaterials such as polysaccharides), and cartilage and/or bone-associatedmaterials (e.g., glucans, dextrans, etc.) as well as microbialmucopolysaccharides. Of course, it is noted that where desirable, suchcompounds may also be synthesized mimetics of the above naturallyoccurring boron and/or borate containing compounds.

Compositions comprising the boron-containing compound may be prepared innumerous manners and may include pharmaceutically and/or nutritionallyacceptable carriers. Therefore, it should be recognized that thecompounds and compositions according to the inventive subject matter canbe modified in numerous manners, and especially preferred modificationsinclude those that improve one or more pharmacokinetic and/orpharmacodynamic parameter. For example, one or more substituents may beadded or replaced at the ligands to achieve a higher AUC in serum. Onthe other hand, and especially where decreased solubility is desired,hydrophobic groups may be added. Moreover, contemplated boron-containingcompounds may be modified to so form a prodrug, and exemplary suitableprotocols for conversion of contemplated compounds into thecorresponding prodrug form can be found in “Prodrugs (Drugs and thePharmaceutical Sciences: a Series of Textbooks and Monographs)” byKenneth B. Sloan (ISBN: 0824786297), and “Hydrolysis in Drug and ProdrugMetabolism: Chemistry, Biochemistry, and Enzymology” by Bernard Testa,Joachim M. Mayer (ISBN: 390639025X), both of which are incorporated byreference herein. On the other hand, and especially where contemplatedcompounds have a higher activity when the compound is metabolized (e.g.,partially hydrolyzed, hydroxylated, glucuronidated, etc.), it should benoted that metabolites of contemplated compounds are also expresslycontemplated herein.

Depending on the particular purpose, it should also be recognized thatcontemplated compounds or combinations of compounds may be combined (invivo or in a pharmaceutical formulation or administration regimen) withfurther pharmaceutically active ingredients, and especially contemplatedother ingredients include known anti-neoplastic, anti-obesity,anti-osteoporosis, anti-stroke, cardioprotective, and/orimmunosuppressant medications.

Depending on the particular use and structure, it is thereforecontemplated that the compounds according to the inventive subjectmatter are present in the composition in an amount generally between 1milligram to 2500 milligrams, more typically between 10 milligrams to1000 milligrams, and most typically between 250 milligrams to 750milligrams per single dosage unit. Thus, preferred concentrations ofcontemplated compounds in vivo or in vitro generally are between 0.1 nMand 500 microM, more typically between 50 nM and 250 microM, and mosttypically between 100 nM and 250 microM. In particularly preferredaspects, and based on various dose range finding studies, especiallypreferred oral dosages are between 200 mg and 1000 mg per day, typicallyin a single dosage (alternatively, up to three daily dosages or evenmore in rare cases, are also deemed suitable). Unless the contextdictates the contrary, all ranges set forth herein should be interpretedas being inclusive of their endpoints, and open-ended ranges should beinterpreted to include only commercially practical values. Similarly,all lists of values should be considered as inclusive of intermediatevalues unless the context indicates the contrary.

Furthermore, it should be recognized that all formulations are deemedsuitable for use herein and especially include oral and parenteralformulations. For example, for oral administration, contemplatedcompositions may be in the form of a tablet, capsule, suspension, orliquid. The nutraceutical or pharmaceutical composition is preferablymade in the form of a dosage unit containing a particular amount of theactive ingredient. Examples of such dosage units are tablets orcapsules. The active ingredient may also be administered by injection asa composition wherein, for example, saline, dextrose or water may beused as a suitable carrier. Alternative suitable routes ofadministration include oral, intramuscular, intravenous, sublingual,rectal, intradermal, transdermal, topical, or subcutaneousadministration. The person of ordinary skill in the art will be wellappraised of appropriate nutraceutical and pharmaceutical carriers, andall of the known carriers are deemed suitable for use herein. Especiallypreferred oral formulations are those in which the boron complex is notretained over a prolonged period, but is bioavailable for absorption inless than 90 after administration. Thus, suitable formulations willinclude those in which contemplated compounds are packaged into softgelatin capsules (with other inactive ingredients as appropriate), orare pressed into a tablet (typically in admixture with pharmaceuticallyacceptable inactive ingredients).

The amount of therapeutically active compound that is administered andthe dosage regimen for treating a disease or condition associated withabnormal levels of calcitriol using compounds and/or compositions ofthis invention depends on a variety of factors, including the age,weight, sex and medical condition of the subject, the severity of thedisease, the route and frequency of administration, and the particularcompound employed, and thus may vary widely. However, especiallysuitable quantities are provided above, and may therefore allow for adaily dose of about 0.01 (or even less) to 100 mg/kg body weight,preferably between about 0.1 and about 50 mg/kg body weight and mostpreferably from about 0.5 to 20 mg/kg body weight. Typically, a dailydose can be administered in one to four doses per day. Especiallycontemplated conditions that though to be modulated with calcitriolinclude neoplastic diseases of the prostate and skin, inflammation,conditions associated with overreaction of the immune system (autoimmunediseases, allergic reactions, etc.), diabetes mellitus (type II),cardiac diseases, and obesity.

For therapeutic or prophylactic purposes, contemplated compounds areordinarily combined with one or more adjuvants appropriate to theindicated route of administration. If administered per os, the compoundsmay be admixed with lactose, sucrose, starch powder, cellulose esters ofalkanoic acids, cellulose alkyl esters, talc, stearic acid, magnesiumstearate, magnesium oxide, sodium and calcium salts of phosphoric andsulfuric acids, gelatin, acacia gum, sodium alginate,polyvinylpyrrolidone, and/or polyvinyl alcohol, and then tableted orencapsulated for convenient administration. Such capsules or tablets maycontain a controlled-release formulation as may be provided in adispersion of active compound in hydroxypropylmethyl cellulose.

Formulations for parenteral administration may be in the form of aqueousor non-aqueous isotonic sterile injection solutions or suspensions.These solutions and suspensions may be prepared from sterile powders orgranules having one or more of the carriers or diluents mentioned foruse in the formulations for oral administration. The compounds may bedissolved in water, polyethylene glycol, propylene glycol, ethanol, cornoil, cottonseed oil, peanut oil, sesame oil, benzyl alcohol, sodiumchloride, and/or various buffers. Other adjuvants and modes ofadministration are well and widely known in the pharmaceutical art.

Of course, it should further be appreciated that all contemplatedcompounds may be present in form of a prodrug, a metabolite, and/or asalt with a pharmaceutically acceptable acid or base. The term “prodrug”as used herein refers to a modification of contemplated compounds,wherein the modified compound exhibits less pharmacological activity (ascompared to the modified compound) and wherein the modified compound isconverted within the body (e.g., in a target cell or target organ) backinto the unmodified form through enzymatic or non-enzymatic reactions.For example, conversion of contemplated compounds into prodrugs may beuseful where the active drug is too toxic for safe systemicadministration, or where the contemplated compound is poorly absorbed bythe digestive tract or other compartment or cell, or where the bodybreaks down the contemplated compound before reaching its target.

With respect to suitable uses, it is generally contemplated that thecompositions and compounds according to the inventive subject matter canbe employed in the prophylaxis and/or treatment of any condition that ischaracterized by an abnormal serum concentration (or kinetic) ofcalcitriol. The term “abnormal serum concentration” as used hereinrefers to a deviation of at least 20% of the average calcitriolconcentration, which is typically below 20 pg/ml, and even moretypically below 15 pg/ml. Therefore, especially contemplated usesinclude all conditions and diseases known to be associated with(permanent or transient) low levels of calcitriol. For example,contemplated uses include prophylactic or therapeutic use ofcontemplated compounds to treat or prevent various cancers, andespecially cancers of the prostate, the mammary gland, and the skin.Moreover, contemplated compounds may also be used to reduceinflammation, or to produce a local or systemic immunomodulation (e.g.,change in Th1/Th2 cytokine balance or immunosuppression). Alternatively,suitable uses further include treatment and prevention of diabetes andheart disease, treatment or prevention of obesity.

EXAMPLES

In a typical example, five subjects were administered a single dosage of552 mg calcium fructoborate upon informed consent, and blood was drawnimmediately prior to administration and after 30, 60, 90, and 120minutes. Calcitriol was measured using a standard radioimmuno assay froma commercial lab starting from 4 ml whole blood (numerous alternativemethods are also deemed suitable, including those described in U.S. Pat.Nos. 4,585,741, 5,202,266, and 6,455,714). From each sample and for eachtime point, concentrations of calcidiol and calcitriol in serum weremeasured and the data are shown in FIGS. 1A to 1D. FIG. 1A depicts theeffect of oral administration of calcium fructoborate on bloodcalcitriol 30 minutes after oral administration, FIG. 1B depicts theeffect of oral administration of calcium fructoborate on bloodcalcitriol 60 minutes after oral administration, and FIG. 1C depicts theeffect of oral administration of calcium fructoborate on bloodcalcitriol 90 minutes after oral administration. FIG. 1D depicts theeffect of oral administration of calcium fructoborate on bloodcalcitriol 90-120 minutes after oral administration. Remarkably, whilecalcitriol serum concentration was significantly and acutely affected ascan be readily taken from FIGS. 1A-1D, the calcidiol concentrationremained unaffected.

FIGS. 2A and 2B depict a graphic illustration of the results from thisstudy where the biphasic mode of modulation of calcitriol is clearlyseen between times 0-90 minutes (decrease) and 90-120 minutes(increase). In most cases, it is expected that the so increasedcalcitriol level will drop back to starting levels within 6 hours orless, and more typically within 4 hours, or less (data not shown). FIG.3 depicts another graphic representation in which the exclusivemodulation of calcitriol versus calcidiol is shown.

As the concentration of calcitriol is tightly regulated, it should beappreciated that the stimulatory effect of contemplated compounds is notexpected to raise the steady-state level of calcitriol, but rather toprovide a pulse of calcitriol (synthesis and/or release). Consequently,contemplated compounds are thought to be particularly useful to providea transient increase in serum calcitriol without significantly alteringserum levels of calcidiol.

While not wishing to be bound by any theory or hypothesis, the inventorscontemplate that the compounds according to the inventive subject matterincrease serum concentration of calcitriol in a bi-modal manner. Duringthe first 60 minutes, a moderate increase is observed in the case ofsubjects with low initial level of calcitriol (<30 pg/ml), but not insubjects with initial level >30pg/ml. Quite likely, activity of the CYPsystem may play a role during this phase. During the 90-120 minuteinterval, all subjects responded the same way with a substantialincrease regardless of initial calcitriol concentration, which mayindicate that the increase in calcitriol may be due to de novo synthesisin the kidney (and possibly ectopically by other tissues and organs).Based on these observations, it should be appreciated that the increaseof calcitriol is not a matter of increased stability of calcidiol aspostulated earlier. Instead, the increase may be due to a stimulatoryeffect of contemplated compounds on the kidney and other enzymaticallyactive tissues. Alternatively, or additionally, stimulation bycontemplated compounds may also be linked to levels in calcium,phosphate, calcitonin, and/or parathyroid hormone.

Notably, it was observed that the serum calcium and serum phosphateconcentrations only changed relatively moderately (about 5-15%) uponadministration of contemplated compounds (and especially administrationof about 550 mg calcium fructoborate), while the serum concentrations ofparathyroid hormone and calcitonin changed at range 10-30% (data notshown).

Among other conditions and disorders that are associated with orresponsive to calcitriol, especially contemplated conditions anddisorders that may be treated with the boron-containing complexespresented herein are those that require reduction of an immune response(e.g., due to activation of monocytes) to so treat certain inflammatoryand/or immune disorders, colorectal cancer, and obesity.

It is still further contemplated that the desirable effect ofselectively modulating calcitriol and not affecting calcidiol may be dueto the relatively high dosage of contemplated compounds. Therefore, itis contemplated that particularly preferred compositions and methodswill include administration of boron-containing compounds at a dailydose of at least 50 mg, more preferably at least 100 mg, even morepreferably at least 250 mg, and most preferably at least 500 mg to aachieve a selective modulation of calcitriol. In further, lesscontemplated aspects of the inventive subject matter, it is noted thatin at least some dosage ranges boric acid and any salt thereof may alsobe employed to modulate the serum calcitriol concentration in a mammal.Thus, it should be appreciated that boron-containing compounds (and tosome extent even boric acid and any salt thereof) may be effective inmodulation of calcitriol and diseases/conditions associated withabnormal calcitriol levels.

Suitable dosage ranges may be identified by various manners, generallyfollowing the experimental protocol as outlined above. Most typically,dosage ranges for a specific route of administration will begin at a lowdose (e.g., 0.1 mg/kg) and increase gradually (typically in two- toten-fold increased of the initial dosage), and effects on bloodcalcitriol will be observed over several hours following administration.Alternatively, suitable dosage ranges can also be determined byreference to specific literature, and even by reference to thisdocument. Similarly, suitable schedules can be determined without undueexperimentation, following the experimental procedures as noted above.Therefore, suitable schedules will typically be between once and fourtimes daily or as otherwise desired.

Using LC-MS, Ca-Fructoborate was detected and measured from serumsamples at concentrations as low as 0.2 ppm and higher. This method wasused to analyze the presence and amount of Ca-Fructoborate in serumcollected from mice treated orally (gavage) with Ca-Fructoborate inliquid at dose 600 mcg/mouse for 30 and 60 minutes. Results show clearlythat Ca-Fructoborate was delivered from intestine to bloodstream inintact form and in time-dependent manner, which is reflected in theTable below. The here obtained data show for the first time thatCa-fructoborate can serve as a controlled-release source of boron inblood and tissues and is not likely to be a pre-from of boric acid.Based upon the amount of Ca-fructoborate provided per mouse (20 g byaverage) the collected results show that bioavailability of the compoundis as high as 556 mcg/ml of serum.

Sample Time of Treatment Intact Ca-fructoborate [mcg/mL] Untreated 0*below detection 1. 30 minutes 449 2. 60 minutes 556 *Detection limit -3 mcg/mL

Detection of intact Ca-Fructoborate in sera collected from fasted micetreated orally with the compound for 30 or 60 minutes. Collected serawere diluted in running buffer and filtered through 0.45 PVDF filter.This solution was analyzed by LCMS for Ca-Fructoborate as thefructoborate anion.

Remarkably, the inventors further contemplate that contemplatedcompounds additionally had a long-term effect on blood/serumtestosterone, particularly where the compounds (typically calciumfructoborate) were administered over extended periods of time. Forexample, upon administration of 100-600 mg of the boron containingcompounds, testosterone levels increased in male and female volunteersat least 15% over base line, more typically at least 20%, and in somecases even more than 25% during administration over at least 14 days.

Consequently, the inventors also contemplate methods of marketing,informing, and advertising a consumer (or patient) that calcitriollevels can be transiently elevated by administration of aboron-containing compound as disclosed and exemplified above. Inespecially preferred methods, the boron-containing compound is acompound in which boron forms a complex with one or more carbohydrate,polyol, and/or amino acid ligands. Furthermore, it is generallypreferred that administration of such compounds is in the range ofbetween 1-2500 mg per day, and more preferably between 100 and 1000 mgper day. With respect to marketing, informing, and advertising, it isnoted that all suitable methods are deemed appropriate and includeprinted matter (e.g., on product package or flyer), displayed matter(e.g., on screen via Internet or TV), and auditory channels (e.g., radioad).

It should be apparent to those skilled in the art that many moremodifications besides those already described are possible withoutdeparting from the inventive concepts herein. The inventive subjectmatter, therefore, is not to be restricted except in the spirit of theappended claims. Moreover, in interpreting both the specification andthe claims, all terms should be interpreted in the broadest possiblemanner consistent with the context. In particular, the terms “comprises”and “comprising” should be interpreted as referring to elements,components, or steps in a non-exclusive manner, indicating that thereferenced elements, components, or steps may be present, or utilized,or combined with other elements, components, or steps that are notexpressly referenced. Where the specification claims refers to at leastone of something selected from the group consisting of A, B, C . . . andN, the text should be interpreted as requiring only one element from thegroup, not A plus N, or B plus N, etc.

1. A method of acutely and transiently increasing endogenous blood calcitriol concentration in a mammal, comprising: identifying a dosage and schedule for administration of a boron-containing compound that is effective to produce a temporary spike in endogenous blood calcitriol concentration in the mammal; and providing the boron-containing compound in a formulation that allows administration of the boron-containing compound in the dosage and schedule effective to produce the temporary spike in the calcitriol concentration in the mammal.
 2. The method of claim 1 wherein administration is oral administration, and wherein the dosage is a daily dosage between 200 mg and 1000 mg of the boron-containing compound.
 3. The method of claim 1 wherein the boron-containing compound is a carbohydrate-boron complex having a boron portion and at least one carbohydrate ligand complexed to the boron portion.
 4. The method of claim 3, wherein the carbohydrate-boron complex has a boron-ligand association constant of between 3,000 and 20,000.
 5. The method of claim 3, wherein the carbohydrate ligand is fructose, mannose, mannitol, sorbose, or sorbitol.
 6. The method of claim 3, wherein the carbohydrate-boron complex is a calcium salt of fructoborate.
 7. The method of claim 3, wherein the carbohydrate-boron complex has a boron-ligand association constant that is sufficient to deliver the carbohydrate-boron complex into blood in measurable quantities.
 8. The method of claim 1, wherein the temporary spike in blood calcitriol concentration is produced by monocyte activation.
 9. The method of claim 1, wherein the administration is effective to modulate an immune response.
 10. The method of claim 1, wherein the immune response is an inflammatory reaction.
 11. The method of claim 1, wherein the temporary spike is an increase of at least 5% of blood calcitriol concentration.
 12. The method of claim 1, wherein the temporary spike is an increase of at least 25% of blood calcitriol concentration.
 13. The method of claim 1, wherein the temporary spike is measurable between 30 and 180 minutes post administration.
 14. A method of modulating a condition associated with a serum calcitriol concentration in a mammal, comprising: identifying a dosage and schedule for administration of a boron-containing compound that is effective to produce a temporary spike in endogenous blood calcitriol concentration in the mammal; and administering the boron-containing compound at the dosage and schedule effective to produce the temporary spike in the calcitriol concentration in the mammal, wherein the temporary spike is effective to modulate the condition.
 15. The method of claim 14, wherein administration is oral administration, and wherein the dosage is a daily dosage between 100 mg and 1000 mg of the boron-containing compound.
 16. The method of claim 14, wherein the boron-containing compound is a carbohydrate-boron complex having a boron portion and at least one carbohydrate ligand complexed to the boron portion.
 17. The method of claim 16, wherein the carbohydrate-boron complex has a boron-ligand association constant of between 3,000 and 20,000.
 18. The method of claim 14, wherein the carbohydrate-boron complex has a boron-ligand association constant that is sufficient to deliver the carbohydrate-boron complex into blood in measurable quantities.
 19. The method of claim 14, wherein the temporary spike in blood calcitriol concentration is produced by monocyte activation.
 20. The method of claim 14, wherein the condition is selected from the group consisting of an inflammatory reaction and a neoplastic condition. 